CN105229139A - For the vessel of cell cultures - Google Patents

Abstract

Have the vessel for cell cultures of the inner compartment of applicable Growth of Cells, in one embodiment, outer tubular wall extends in a longitudinal direction and delimit the outer edge of described inner compartment in radial directions.First outer end and the second outer end respectively at the first outer end of outer tubular wall and the second outer end place to described inner compartment gauge.Fixing weighting material in described inner compartment comprises the weighting material of such as fibre substrate.Also disclose other embodiment and relevant method.

Description

For the vessel of cell cultures

The cross reference of related application

This application claims U.S. Provisional Patent Application No.61/735, the right of priority of 841, its disclosure is incorporated to by way of reference at this.

Technical field

The disclosure relate to for cell cultures vessel, comprise the vessel for cell cultures bio-reactor, manufacture the method for this vessel or bio-reactor and use the method for these vessel or bio-reactor, such as culturing cell.

Background technology

In cell cultures now, the output comprising the bio-reactor of one or more vessel is one of multiple principal concern of biological industry.The complicacy of current method also may raise the cost.Thus, determine that needs improve.

Summary of the invention

An object of embodiment disclosed herein is to provide the excellent vessel for cell cultures and method.Its advantage is, when for Growth of Cells, the output of described vessel can at least equal or even be greater than the output of current known vessel, and can not increase complicacy or cost.An advantage is to provide the vessel with more high-cell density, thus provides " volumetric production " of increase, i.e. the output of every vessel volume.Advantageously, the vessel can when being used for Growth of Cells with cell cultures basically identical in whole vessel are provided.Advantageously provide following vessel: when for Growth of Cells, have along the whole surface that cell grows evenly medium ventilation and upgrade.Advantageously provide following vessel: these vessel provide the viability of good Growth of Cells and/or improvement to safeguard.Advantageously providing can for the manufacture of the vessel of disposable vessel and bio-reactor.

It is one or more that a kind of method and vessel can realize in above-mentioned purpose.

According to first aspect, the embodiment according to the vessel for cell cultures of the present disclosure has internal space, and this internal space comprises the weighting material of applicable Growth of Cells.This internal space is limited by following parts:

-outer tubular wall, this outer tubular wall has the first outer end and the second outer end,

-Nei extends wall, and the first outer end of this interior elongation wall is directed towards the first outer end of outer tubular wall, and the second outer end of this interior elongation wall is directed towards the second outer end of outer tubular wall, and this interior elongation wall is positioned at outer tubular wall, and

-the first capping and the second capping, this first capping and the second capping are used for closing described internal space.

Vessel for cell cultures can have annular internal space, and this annular internal space comprises the weighting material of applicable Growth of Cells.When it is present, this annular internal space has by the ring-type volume limited with lower component:

-outer tubular wall, longitudinal wall that this outer tubular wall has the first outer end and the second outer end and extends in a longitudinal direction, this outer tubular wall delimit the outer edge of described ring-type volume in a longitudinal direction;

-the first capping and the second capping, this first capping and the second capping are carried out gauge in the first outer end of outer tubular wall and the second outer end place to described ring-type volume and are closed respectively;

-Nei extends wall, first outer end of this interior elongation wall is directed towards the first outer end of outer tubular wall, second outer end of this interior elongation wall is directed towards the second outer end of outer tubular wall, this interior elongation wall is positioned at outer tubular wall, this interior elongation wall extends in a longitudinal direction and delimit the inner boundary of described ring-type volume, and this inner boundary is surrounded by described outer edge.

Alternatively, this inner boundary is by described outer sealed boundary.

Can align with the first capping in first outer end of this interior elongation wall.Can align with the second capping in second outer end of this interior elongation wall.

This interior elongation wall can be provided by interior elongate member.This outer tubular wall can be provided by outer tubular member.

Advantageously, can force to be arranged in described vessel and flow through described weighting material with the developing medium (such as cell culture medium) of the basic all volumes allowing cell cultures.

According to an embodiment, tangent and the smallest circle cylinder with axis in a longitudinal direction can have diameter Dmax with described outer edge; Distance radially between described inner boundary and outer edge can in the scope of 5% to 45% of Dmax.

The annular internal space with ring-type volume has following advantages, that is: during Growth of Cells, once rotate the vessel being filled with developing medium (such as cell culture medium) at least partly, medium just can flow through described weighting material with substantially homogeneous and/or constant speed.This medium even can flow through described weighting material with the form of plug flow.Can obtain evenly and predictable Growth of Cells.Alternatively, higher output is obtained.When medium being mixed with gas (such as oxygen or carbonic acid gas) to allow Growth of Cells, medium via the measurable and more homogeneous flowing of described weighting material can cause to described weighting material evenly ventilation, therefore can improve the Growth of Cells efficiency on whole weighting material.It is especially true when developing medium (such as cell culture medium) flows through described weighting material as plug flow.

When developing medium (such as cell culture medium) penetrates in the given area of described weighting material, force existing gas or air displacement, such as, flow out this region.Once the developing medium of certain volume (such as cell culture medium) is through this region, just produce low pressure at developing medium (such as cell culture medium) rear of discharged certain volume.This low pressure causes the gas that exists in adjacent weighting material region or air to be inhaled into existing in the region of low pressure.Like this, not only developing medium (such as cell culture medium) is shifted through described weighting material, and the gas existed in these vessel or air are also through described weighting material displacement.

Developing medium (component of such as cell culture medium, particularly this medium) can be suitable for obtained output.In a particular embodiment, this medium is rich in such as spore element or the other carbon source of of short duration or constitutive expression.In other specific embodiment, such as, employ minimum medium in order to labelling experiment (such as NMR mark).

According to the part that the vessel of first aspect can be the bio-reactors for cell cultures.These vessel can rotatably be arranged in bio-reactor, and thus, these vessel rotate around an axis, and this axis is basically parallel to the longitudinal direction of described vessel, i.e. the longitudinal direction of outer tubular wall.Two outer ends place being sealed on outer tubular wall closes its ring-type volume.This prevent the medium that exists in this volume these cappings At The Height transversely direction from this ring-type volumetric leak.

In any embodiment, this weighting material can be the weighting material of applicable adherent cell growth.This weighting material can be the weighting material of applicable suspension culture.The group of the optional freedom of this weighting material woven or nonwoven primitive fiber, porous carbon and glycan substrate composition.Alternatively, this primitive fiber can be made up of PET or other polymkeric substance any or biological polymer.Polymer treatment can be become be applicable to cell cultures, if this process is necessary.

The weighting material be applicable to or " solid support material " are the mineral carrier of the such as silicate of applicable Growth of Cells, the natural product of the organic compound of calcium phosphate, such as porous carbon, such as chitosan, polymkeric substance or biological polymer.This weighting material can adopt the form of the pearl with rule or erratic composition, or can comprise the polymkeric substance of applicable Growth of Cells or the woven of other material any or nonwoven primitive fiber.Also this weighting material can be set to single with multiple hole and/or passage.Weighting material in these vessel can have various forms and size.In certain embodiments, this weighting material is solid or porous ball, thin slice, polygonal particulate material.Usually, use enough weighting materials to move in vessel in use to avoid this filler particle, because this may damaging cells and can affect the circulation of gas and/or medium.Alternatively, this weighting material by being coupled to inner vessel therein or the element be coupled in the compartment of vessel is formed, and has suitable porosity and surface.According to specific embodiment, the scope of the porosity of this weighting material is 60% to 99%, more particularly, is 80% to 90%.In order to adherent cell growth, the cell of this weighting material can reach surface about 150 to 1000cm ²/ cm ³between.An one example is the carbon (Carboscale) that Cinvention company (Germany) manufactures.

Can be the outer tubular wall of these vessel, interior elongate member, capping and optionally other element provide these vessel applicable to carry out the material of cell cultures, such as stainless steel, polystyrene, glass, polyethylene, methyl methacrylate, high density polyethylene(HDPE), Low Density Polyethylene, polyethylene terephthalate glycol, perfluoroalkoxy resin, polycarbonate, poly(vinylidene fluoride), tetrafluoroethylene, ultrahigh molecular weight polyethylene(UHMWPE), nylon, special teflon, crystal polystyrene, metallocene polypropylene or syndiotactic polystyrene.In one embodiment, wherein, cell suspension grows, as required with the element of these vessel of compound treatment preventing cell adhesion.In another embodiment, wherein, cell grows as adherent cell, uses the element of these vessel of compound treatment of such as polylysin as required, to obtain sticking of expectation.

These vessel also can comprise at least the first and second fluid penetrable partition members, and described internal space is divided at least two compartments by this first and second fluid penetrable partition member, wherein:

-this fluid penetrable partition member extends to outer tubular wall from interior elongation wall and extends to the second capping from the first capping on the longitudinal direction parallel with the direction of tubulose axis, and

One in-described first compartment and the second compartment does not comprise weighting material.

These vessel also can comprise at least N number of fluid penetrable partition member, and described internal space is divided at least N number of compartment by them, N be equal to or greater than 2 integer.These fluid penetrable partition members can extend to outer tubular wall from interior elongation wall and extend to the second capping from the first capping on the longitudinal direction parallel with the direction of tubulose axis.Alternatively, maximum N-1 compartment is provided with described weighting material.Alternatively, N is even number, and N/2 compartment is provided with described weighting material.When N is equal to or greater than 2, each compartment comprising described weighting material can adjoin with two of a non-filler compartment.

When outer tubular wall is cylindrical wall, described internal space is divided into the multiple compartments extended on the direction that the direction of the longitudinal direction with outer tubular wall is parallel.

The volume of the compartment of non-filler can between 25% to 90% of the ring-type volume of described internal space.

This fluid penetrable partition member can transflective liquid and gas, and can be applicable to keeping described weighting material.

This partition member can be made of porous materials, and its multiple hole body has and is applicable to making partition member to the permeable size of liquids and gases, and these hole bodies are enough little, so that this partition member can fix described weighting material.

As an alternative, this partition member is for also can be saturating used liquids and gases.Like this, open spaces can be divided into multiple subelement, and wherein each subelement can be provided with suitable weighting material.

At least one in first capping and the second capping can be provided with at least one junctor, provides medium and/or gas for internally space and/or extracts medium and/or gas from internal space.

This junctor can comprise snap-fitting or the joint that closely cooperates, or can be other suitable junctor any.This junctor or multiple junctor can be connected to flexible conduit or pipe or pipeline section, and medium is guided to internal space or draws from internal space by this flexible conduit or pipe or pipeline section.In a particular embodiment, the joint that closely cooperates is connected to flexible duct.

The component of medium depends on the type of considered cell.These media be those skilled in the art known, and can commercially to buy to each manufacturers.Consider the high-cell density adopting the vessel of embodiments of the invention to obtain, in a particular embodiment, this medium is rich in carbon source, spore element, VITAMIN, amino acid etc.In a particular embodiment, also to this medium supply schedule surface-active agent, to adapt to the rheological property of this medium.Described gas normally air, for the growth of bacterium and yeast, and mammalian cell usually 95% air and 5%CO ₂mixture in grow.In ad hoc approach embodiment of the present invention, monitor oxygen and/or the CO of the gas in these vessel ₂loss, and change or supplemental oxygen or CO ₂.

At least one in first capping and the second capping can be provided with at least one junctor, provides medium and/or gas with internally space and/or from internal space, extracts medium and/or gas.This at least one capping can comprise this at least one junctor, and it is positioned such that to provide medium and/or gas and/or extract medium and/or gas from this compartment in the compartment that this at least one junctor is applicable to not existing wherein weighting material.At least one in first capping and the second capping can comprise a more than junctor, provides medium and/or gas with in internally space and/or from internal space, extracts medium and/or gas.

In at least one and the separable situation of outer tubular wall in the first capping comprising this at least one junctor and the second capping, this at least one junctor be applicable to being positioned such that this at least one junctor for there is not weighting material wherein compartment in medium and/or gas are provided and/or extract medium and/or gas from this compartments.

This at least one junctor can comprise the joint that closely cooperates for connecting flexible conduit.

This at least one junctor can be arranged in described capping near outer tubular wall.

Described opening can be placed in contact outer tubular wall.

Described interior elongate member can be positioned at the center of outer tubular wall.

Described outer tubular wall can be provided by cylindrical tubular members.

Described interior elongation wall can be made up of cylindrical elements.

Interior elongate member can be disposed coaxially in outer tubular wall.

Described interior elongate member can have the radial section based on the plane vertical with its longitudinal direction.This radial section can have the shape selected from following group: circle, oval, ellipse, square, rectangle and circle are arc.

This radial section can have circular segment shape, and this circular segment has the central angle being greater than 180 °.This embodiment has following advantages, i.e.: when these vessel rotate about the axis (this axis is the axis of outer tubular member and interior elongate member alternatively), can obtain the position of rotation producing gas chamber.When these vessel rotate to wherein above-mentioned string basic horizontal and described circular segment is positioned at the position above this string, this gas chamber can be obtained.When these vessel be filled make the liquid level of medium lower than this string time, the space between medium liquid level and this string can be used as gas chamber.Exchanging gas between the outside outside of bio-reactor (alternatively for) that free dielectric surface is adapted at these vessel (being a part for bio-reactor alternatively) and these vessel.This chamber can be used to make it enter or flow out these vessel gas will not with physical contact between media.

Described interior elongation wall can be tubular wall.

Described interior elongation wall can be connected in the fixed position for outer tubular wall.

Described interior elongate member can be provided by interior elongate member, and described outer tubular member is provided by outer tubular member, and this interior elongate member and outer tubular member are by shaping integratedly.

Described interior elongate member and outer tubular member can relative to each other rotate.

Described outer tubular wall can be cylindrical tubular wall, and it has the radial section based on the plane vertical with its longitudinal direction, and this radial section has the shape selected from following group: circle, oval, ellipse, square or rectangle.

Described outer tubular wall and interior elongation wall can be cylindrical tubular wall.The radial width of the described internal space between described outer tubular wall and interior elongation wall can in the scope between 5% to 45% of the diameter of outer tubular member.

Described interior elongation wall can be disposed coaxially in outer tubular wall.

Described internal space can have 100 milliliters to 25 liters or the even volume of 100 liters.

The diameter of described outer tubular wall and the length ratio of vessel can between 10/25 to 100/25.Therefore, the diameter of the outer tubular member of these vessel can be 0.4 to 4 times of the length of vessel.

At least one comprised connection element in first capping and the second capping, for being attached to the driver element for rotating this outer tubular wall by outer tubular wall.

Described interior elongation wall can be provided by interior hollow elongated element, the first outer end of this interior elongate member of the first lidstock.

Above-mentioned interior elongation wall can be provided by interior hollow elongated element, the second outer end of this interior elongate member of the second lidstock.

Described interior elongation wall can be fluid penetrable wall.According to some embodiments, described outer tubular wall can be fluid penetrable wall.According to some embodiments, described interior elongation wall and/or outer tubular wall can transflective liquid or gases, and are applicable to keeping described weighting material.

The porous material comprising multiple hole body can be used to provide described fluid penetrable wall, therefore provide porous wall.As an alternative, the perforated material comprising multiple hole can be used to provide fluid penetrable wall, therefore provide perforated wall.But described fluid penetrable wall can be provided with enough little hole body or hole, wear to opposite side from the side of this wall to prevent described weighting material.

According to second aspect, provide a kind of bio-reactor for cell cultures.This bio-reactor comprises the one or more vessel according to disclosure first aspect.These one or more vessel can be arranged so that their tubulose axis basic horizontal and can longitudinally rotate by axis.

The longitudinal axis of these vessel is axis parallel with the longitudinal direction of outer tubular wall.These one or more vessel can rotatably be arranged in reactor vessel.Alternatively, outer tubular wall is provided with multiple hole body, or is made up of porous material (such as liquids and gases permeable material).When fluid penetrable outer tubular wall rotate in the container being partially filled medium make only outer tubular wall a part of submergence in media as well time, can be described weighting material and the medium flowed into through outer tubular wall in ring-type volume be provided.When these vessel rotate in container, a part for medium can be pulled together with described weighting material.This liquid permeable material is used to also simplify the control of the method.The vessel closed require to use junctor and carry out the chemical examination of medium and gas or supplement.This requires suitably this junctor of location, and the time suitably selected sampling or supplement.One or more vessel wherein with fluid penetrable outer wall, by the embodiment that is rotatably placed in more large container, can perform the sampling to gas and medium constantly in this container.

These one or more vessel can comprise flexible conduit, for the internal space of these vessel being attached to the holder of medium and/or gas.

Described bio-reactor can comprise drive unit, and this drive unit alternately can rotate described outer tubular wall clockwise and counterclockwise.

Alternatively, described interior elongation wall and outer tubular wall synchronous rotary.

This bio-reactor also can comprise driving roll, carrys out spun outer tube shape wall for the outside surface by contacting outer tubular wall.

Described vessel can comprise magnetics.This bio-reactor also can comprise magnetics, and this magnetics cooperates with the magnetics of described vessel.These two magneticss can be located so that the magnetics of this bio-reactor will cause the rotation of the magnetics of described vessel around the rotation of rotation, therefore by making the outer tubular wall of described vessel rotate, alternatively, whole vessel are rotated.

According on the other hand, provide a kind of method for culturing cell.The method comprises the following steps:

A) following material is incorporated in vessel according to a first aspect of the present invention:

-attached cell or non-adherent cell, and

-developing medium,

B) described vessel are located so that the longitudinal direction basic horizontal of outer tubular wall, this outer tubular wall can rotate along the axis parallel with its longitudinal direction, and

C) outer tubular wall of described vessel is rotated along described axis.

Alternatively, described interior elongation wall can with outer tubular wall synchronous rotary.

According to some embodiments, obtain the cultivation of cell by adherent cell growth.

According to some embodiments, the cultivation of cell can be obtained by suspension culture.

According to some embodiments, the upper flow liquid level of described developing medium can at least reach described interior elongation wall when making vessel be at least one radial position.

Upper flow liquid level (being liquid levels in this case) can be arranged in internal space, so that when being at least one radial position, this upper flow liquid level is located to contact described interior elongation wall at least on one point.Described interior elongation wall can partly submergence be in media as well when being at least one radial position.

According to some embodiments, the upper flow liquid level of described developing medium can when vessel are at least one radial position lower than described interior elongation wall.

Upper flow liquid level (being liquid levels in this case) can be arranged in internal space, so that when being at least one radial position, this upper flow liquid level is not located to extend wall in contact on one point.This provides fluid surface freely, on this fluid surface, there is gas chamber.This gas chamber can be used for exchanging from the gas of this vessel outside or by the outside of gaseous interchange to vessel.

According to some embodiments, the volume of the medium arranged in open spaces can between 25% to 100% of the ring-type volume of the internal space of these vessel.

According to some embodiments, described outer tubular wall alternately clockwise and be rotated counterclockwise.

According to some embodiments, can with the rotation of the maximum angular rate execution between per minute 0.1 to 25 turn around described axis.

According to some embodiments, described tubular wall can have and is directed the outside surface contrary with internal space.This outside surface can have the maximum line velocity being up to 10cm/ second.

According to some embodiments, the method can comprise the step of chemical examination and/or the medium changed in the internal space of these vessel and/or gas.

According to further aspect of the present disclosure, have for Growth of Cells inner compartment, comprise for the vessel of cell cultures:

-outer tubular wall, this outer tubular wall extends in a longitudinal direction, and this outer tubular wall delimit the outer edge of described inner compartment in radial directions;

-first end and the second end, this first end and the second end respectively at the first end of outer tubular wall and the second end place to described inner compartment gauge; With

-weighting material, this weighting material is in described inner compartment and comprise fibre substrate.

In one embodiment, described first end and the second end comprise removable lid.This first end can comprise entrance, and the second end can comprise outlet.Can one or more fluid penetrable dividing plate be set in described compartment, for contacting this fibre substrate.

Can provide at least for rotating the turner of described weighting material.This turner can comprise the roller for contacting with the outer tubular wall of vessel.This turner can comprise magnetic and connect.

A closed loop can be provided, for delivering the fluid to described compartment and transferring out from described compartment.Reservoir can be connected to this closed loop, and at least one sensor is associated with this reservoir.The group of the following item composition of the optional freedom of this sensor: temperature sensor, position transducer, optical pickocff, pH sensor, oxygen sensor, CO ₂the combination of sensor, ammoniacal sensor, cell density sensor or any the sensor.

In any embodiment, the fiber of described fibre substrate can comprise nonwoven PET.

Further aspect of the present disclosure relates to a kind of for making the equipment of Growth of Cells, comprising:

-roller bottle, this roller bottle comprises: outer tubular wall, and this outer tubular wall extends in a longitudinal direction; First end and the second end, this first end and the second end respectively at the first outer end of outer tubular wall and the second outer end place to described compartment gauge; And weighting material, this weighting material is positioned at described compartment and comprises fibre substrate; With

-turner, this turner is for rotating described weighting material.

In one embodiment, the first and second isolated first fluid permeable separator are positioned at described compartment.A closed loop can be provided, for delivering the fluid to described compartment and transferring out from described compartment.Reservoir can be connected to this closed loop, and at least one sensor is associated with this reservoir.

Further aspect of the present disclosure relates to a kind of vessel for cell cultures, this container has the inner compartment for Growth of Cells, these vessel comprise: the outer tubular wall extended in a longitudinal direction, and this outer tubular wall delimit the outer edge of described inner compartment in a longitudinal direction; With first end and the second end, this first end and the second end respectively at the first outer end of outer tubular wall and the second outer end place to described compartment gauge; Be in the weighting material in described compartment; With at least one the fluid penetrable dividing plate being positioned at described compartment.In one embodiment, this dividing plate comprises perforation plate.

Another aspect of the present disclosure relates to a kind of method for culturing cell, comprising:

The roller bottle with weighting material is provided;

Transport fluid in roller bottle; With

Rotate this weighting material.

In one embodiment, the step rotating this weighting material (this weighting material can comprise fibre substrate) rotates this roller bottle after being included in described supplying step.The step rotating this weighting material can be included in roller bottle and keep fixed and rotate this weighting material simultaneously.During the step of extracting fluid from roller bottle out can occur in this spin step.The method also can be included in the step of extracting test fluid characteristic after step out, and makes extracted out fluid be back to step in roller bottle during being included in described supplying step further.

Further aspect of the present disclosure relates to a kind of vessel for cell cultures, and this container has the inner compartment for cell cultures, comprising:

-the outer tubular wall that extends in a longitudinal direction, this outer tubular wall delimit the outer edge of described inner compartment in a longitudinal direction;

-first end and the second end, this first end and the second end respectively at the first outer end of outer tubular wall and the second outer end place to described compartment gauge;

-be in weighting material in described compartment; With

-conduit, the fluid being used for culturing cell is transported in inner compartment by this conduit.

In one embodiment, this conduit comprises flexible tubing.These vessel also can comprise for being connected to the entrance of this flexible tubing and being connected to the outlet of this flexible tubing.This conduit can comprise the first flexible tubing being connected to entrance and the second flexible tubing being connected to outlet.This entrance and exit can be arranged on the common wall of described vessel.This entrance can be connected to the pipe extended in described inner compartment, with away from outlet position conveyance fluid.Sensor can be connected to these vessel, and described sensor is associated with flexible hose line.Insufflator (such as spinner-type flowmeter) can be connected to this conduit.Can swivel joint be provided with, with this tubes connection.

In any embodiment, the system for culturing cell all can comprise the vessel being positioned at and comprising on the recirculation loop of this conduit.This conduit can be connected to described vessel at one end, and is connected to reservoir at the other end place.This reservoir can comprise strainer and/or sensor.

Of the present disclosurely relate in one aspect to again a kind of method for culturing cell, comprising:

A () rotates the container comprising fixing weighting material along first direction; With

B (), after rotating along first direction, rotates this container along second direction.

The method delivers the fluid to container during can being included in the step rotated along first direction.The method delivers the fluid to container during can being included in the step rotated along first direction.Container to be connected to the step of the conduit comprising flexible tubing by the method before also can being included in supplying step.This fluid can comprise liquid, and the method also can be included in supplying step before by gas injection to the step in liquid.

In any embodiment, all can complete this rotation to the complete at least one times rotation on each direction of container in first direction and second direction.This rotation can be completed at least twice complete rotation on each direction of container in first direction and second direction.This rotation can be completed at least three complete rotations on each direction of container in first direction and second direction.The method also can comprise repetition above-mentioned steps (a) and (b).

Specific and preferred aspect is proposed in appended independent sum dependent claims.Optionally, the feature of dependent claims can with the feature of independent claim and combined with the feature of other dependent claims, and be not only clearly propose in the claims.

Although the device of this area has existed lasting improvement, change and change, believe that this design represents the improvement of novelty in fact, comprised and the departing from of former practice, thus provided the more efficient, stable of this character and device reliably.

This instruction allows to use suspension culture or adherent cell growth to improve the design of the bio-reactor for cell cultures.Described vessel can be used for the prokaryotic cell prokaryocyte of such as such as bacterial cell, particularly eukaryotic cell such as yeast cell, vegetable cell, insect cell, Mammals and particularly human cell, or produce the antibody of hybridoma or stem cell.According to an embodiment, these cells itself are gathered in the crops.In other embodiments, have collected cell-derived product, such as, from virus, antibody, recombinant nucleic acid structure (such as, plastid) that infected cell obtains, recombinant protein or cell metabolite, such as VITAMIN or hormone.

From the detailed description provided below in conjunction with accompanying drawing, above-mentioned and other characteristic, feature and advantage will become more obvious, and these accompanying drawings show principle of the present invention as an example.This explanation only provides as an example, not limits the scope of the invention.The reference drawing hereafter quoted refers to accompanying drawing.

Accompanying drawing explanation

Fig. 1 is the longitudinal section view of schematic side elevation according to the vessel of disclosure first aspect and these vessel, and the longitudinal section view in Fig. 1 completes based on the plane of the central axis upright with vessel.

Fig. 2 is the schematic diagram of the longitdinal cross-section diagram of vessel according to disclosure first aspect, and this sectional view is based on the centerline axis parallel with vessel and the fore-and-aft plane comprising this central axis completes.

Fig. 3 is side-view according to the vessel of first aspect and longitudinal section view, and the cross section of Fig. 3 a completes based on the plane of the central axis upright with vessel.

Fig. 4 a-4c is the side-view according to the longitudinal section view of the alternative vessel of first aspect and the circular segment as a vessel part, and this sectional view completes based on the plane of the central axis upright with vessel.

Fig. 5 a and Fig. 5 b is the longitudinal section view of the bio-reactor of the vessel comprised according to first aspect, and this sectional view completes based on the plane of the central axis upright with vessel.

Fig. 6 is the longitdinal cross-section diagram of the bio-reactor comprising multiple vessel according to first aspect, and the sectional view of Fig. 6 a is based on the centerline axis parallel with vessel and the fore-and-aft plane comprising this central axis completes.

Fig. 7 diagrammatically illustrates cross-sectional view and the longitudinal cross-section of the vessel be as shown in Figure 3 arranged on cooperation drive shaft.

Fig. 8 a-8c diagrammatically illustrate be arranged on cooperation drive shaft, according to the alternative vessel of embodiment.

Fig. 9 diagrammatically illustrates the longitudinal section view of the bio-reactor comprising vessel, and these vessel are according to an embodiment of the present disclosure.

Figure 10 and 10a-10c shows alternate embodiments.

Figure 11-19 shows further aspect of the present disclosure.

In various figures, identical Reference numeral refers to same or analogous element.

Embodiment

The disclosure comprises the explanation of specific embodiment and with reference to certain figures, but the present invention is not limited thereto, but is only defined by the claims.Described accompanying drawing is only schematic and nonrestrictive.In the accompanying drawings, the size of some elements may be exaggerated in order to illustrate, and can not to scale (NTS) draw.These sizes and relative dimension also do not correspond to the actual reduction of the present invention's practice.

In addition, term " first ", " second ", " the 3rd " etc. in specification sheets and claim for distinguishing between like, and not necessarily is with time, space, grade or describe a kind of order in any other way.Should be appreciated that the term so used can exchange in appropriate instances, and embodiments of the invention as herein described can with other sequential operation different from described herein or shown order.

In addition, the term " top " in specification sheets and claim, " bottom ", " on ", " under " etc. are only for descriptive object, and not necessarily describes its relative position.Should be appreciated that the term so used can exchange under suitable situation, and embodiments of the invention as herein described can with other direction operation different from described herein or shown direction.

It should be noted that not should by the term used in claim " comprising ... " be interpreted as being limited to the meaning listed hereinafter; It does not get rid of other element or step.Thus, should be interpreted as showing described feature, entirety, step or the parts involved by existing, but not get rid of and also there are or add one or more further feature, entirety, step or parts, or their group.Thus, the scope of " device comprises component A and B " stated should not be limited to this device and only be made up of components A and B.It only means: about the disclosure, and unique associated components of this device is A and B.

Similarly, should understand, the term yet used in the claims " connection " should be interpreted as being only limitted to direct connection.Term " connection " and " connection " and their derivative words can be used.Should be understood that these terms are synchronized with each other unintentionally.Thus, the output terminal that the scope of " device A is attached to device B " stated should not be limited to wherein device A is connected directly to device and the system of the input terminus of device B.It is meant between the output terminal and the input terminus of B of A, there is path, this path may be the path comprising other device or component.The meaning of " connection " can be two or more element direct physical or electrical contacts, or two or more elements directly do not contact but still coordination with one another or interaction each other.

" embodiment " mentioned in whole specification sheets or " embodiment " refer to the special characteristic, structure or the characteristic that comprise at least one embodiment and describing in conjunction with this embodiment.Thus, word " in one embodiment " or " in one embodiment " of each local appearance in whole specification sheets must not relate to same embodiment, but likely relate to same embodiment.In addition, according to the disclosure, those skilled in the art it should be understood that described special characteristic, structure or characteristic can combine in one or more embodiments in any suitable manner.

Similarly, should understand, in the explanation of exemplary embodiment, in order to make the disclosure smooth and one or more in order to what help to understand in each inventive aspect, sometimes by various feature of the present invention single embodiment, accompanying drawing or its illustrate in combine.But, this method of the present disclosure should be interpreted as show that claim needs the more feature than clearly listing in each claim.On the contrary, as claims reflect, each inventive aspect is less than all features of single disclosed embodiment above.Thus, the claim after detailed description should be incorporated in this detailed description clearly, every bar claim itself is all as an independent embodiment of the present invention.

In addition, although embodiments more as herein described comprise some but not further feature that other claim comprises, it will be understood by those skilled in the art that the characteristics combination of different embodiment is had a mind within the scope of the invention, and form different embodiment.Such as, in the following claims, any embodiment required for protection may be combinationally used with any.

In addition, the element as herein described of apparatus embodiments is an example of the component in order to realize the function that the present invention is performed by this element.

Many specific detail have been set forth in explanation provided herein.However, it should be understood that and can put into practice embodiments of the invention when there is no these specific detail.In other cases, in order to not hinder the understanding to this explanation, not shown well-known method, structure and technology.

Only provide following term to help to understand the present invention.

" radial direction " is interpreted as the axes normal with minimum circumcircle cylinder.

" circumscribed " is interpreted as being configured to, such as, constructing or be built into around a geometricdrawing to contact point as much as possible.

Term " tubulose " is interpreted as hollow volume, extends in a longitudinal direction.

Term " cylindrical " is interpreted as having cylindrical shape." right cylinder " is interpreted as following objects: its have by along and fixing straight line (i.e. longitudinal direction) extend abreast and with fix or surface that the track of straight line that closed curve intersects is formed and the volume that two parallel planess cutting this surface limit.

Circular segment or " cutting circle " provide one of two parts obtained during two point of crossing when circle and straight line intersection.In every part, be all called string by along this straight line from the path in the first point of crossing, point of crossing to the second.For every part, all there is the path along this circle from the first point of crossing, point of crossing to the second, it is arc section.For every part, all the first point of crossing will be rotated the central angle being called circular segment with the angle making the first point of crossing overlap with the second point of crossing along this arc section.The height of this circular segment is the distance between the string at half central angle place and arc section.It is arc radius of a circle that this height can be greater than its shape.The surface of this circular segment can be greater than the half that its shape represents the surface of arc circle.

Now inventive concept disclosed herein is described the detailed description by several embodiment.Obviously, when not departing from true spirit of the present invention or technology instruction, can construct other embodiments of the invention according to the knowledge of those skilled in the art, the present invention only limits by the term of claims.

First embodiment of the vessel 1 according to disclosure first aspect has been shown in Fig. 1, and these vessel 1 are suitable for use in bio-reactor.Vessel 1 for cell cultures have internal space 2, and this internal space 2 can be ring-type as shown in the figure, but also can take other form as herein described.Space 2 comprises weighting material 3.When these vessel are used for cell cultures, weighting material 3 should be applicable to Growth of Cells.

In shown atoll texture, internal space 2 has by with the ring-type volume of lower component gauge:

-outer tubular wall 4, longitudinal wall 43 that this outer tubular wall 4 has the first outer end 41 and the second outer end 42 and extends on the longitudinal direction shown in arrow 44.Outer tubular wall 4 delimit the outer edge of this ring-type volume on longitudinal direction 44;

-the first capping and the second capping (61,62), this first capping and the second capping (61,62) also close this ring-type volume in the first outer end 41 of outer tubular wall 4 and the second outer end 42 place to described ring-type volume gauge respectively;

-Nei extends wall 5, and the first outer end 51 of this interior elongation wall 5 is directed towards the first outer end of outer tubular wall, and the second outer end 52 of this interior elongation wall 5 is directed towards the second outer end of outer tubular wall 4.Interior elongation wall 5 is positioned at outer tubular wall 4.Interior elongation wall 5 extends and delimit the inner boundary of described ring-type volume on longitudinal direction 44, and this inner boundary is surrounded by described outer edge.

Align with the second capping 62 in second outer end 52 of interior elongation wall 5.As an example, outer tubular wall is provided by cylindrical outer tube linear element 40.Interior elongation wall 5 can by solid interior cylindrical elements 50, such as cylindrical bar.Outer tubular member 40 is cylindrical tubular members, and has the central axis 45 being parallel to longitudinal direction.Interior cylindrical elements 50 and outer tubular member 40 can be installed coaxially.

First outer end of interior cylindrical elements 50 can comprise connection element 53 so that interior cylindrical elements 50 is attached to driving mechanism, and this connection element 53 is also by being attached to driving mechanism with interior cylindrical elements 50 and the fixing capping 61 and 62 of outer tubular member 40 by outer tubular member 40, and this driving mechanism is such as the motor of bio-reactor.Second capping 62 is provided with junctor, and this junctor is applicable to described vessel to be attached to medium source or gas source, provides medium and/or gas and or extract medium and/or gas from internal space 2 with internally space 2.Can be the first capping 61 or the second capping 62 provides this junctor or other alternative junctor.

Internal space 2 is filled with weighting material 3 at least in part.As an example, this weighting material can be the mineral carrier of the such as silicate of applicable Growth of Cells, the natural product of the organic compound of calcium phosphate, such as porous carbon, such as chitosan, polymkeric substance or biological polymer.This weighting material can have and adopts rule or the bead form of erratic composition, or can comprise the polymkeric substance of applicable Growth of Cells or the woven of other material any or nonwoven primitive fiber.Also this weighting material can be set to single with multiple hole and/or passage.

Alternatively, this weighting material can have the porosity P in 50% to 98% scope.Term " porosity P " is the volume of air existed in the material of given volume, and is represented as the per-cent of the material of given volume.Weight Wx by each volume measuring porous material measures porosity, and uses following formula:

P＝100-(1-Wx/Wspec)

Wherein, Wspec is the proportion of material.This porous material can be the porous material unit of a solid, or can be multiple independent unit, such as particle, fragment, pearl, fiber or fiber block.

When mobile described vessel, this weighting material (particularly porous material) can be in fixing relative position relative to described vessel, or can move relative to described vessel in described vessel, or can move in the compartment of described vessel depending on particular case.

Described vessel can rotate around its axis 45, alternatively, rotate with the rotating speed of per minute 0.1 to 25 turn.

As shown in the best in the longitudinal section view of Fig. 1 a, this internal space is filled with developing medium partly, such as cell culture medium 15.As an example, extend wall 5 in liquid level 18 at least contacts, or as shown in the embodiment of figure 1, interior elongation wall 5 is partially immersed in medium 15.The part be positioned under liquid level 18 of weighting material 3 is soaked by developing medium (such as cell culture medium).The weighting material 3 be positioned on liquid level 18 contacts gas or the air of internal space 2 existence.When described vessel rotate around axis 45 in one direction, such as shown in arrow 17 turn clockwise time, developing medium (such as cell culture medium 15) is with regard to reverse rotation, and namely relative to weighting material 3, with counterclockwise, (shown in 19) rotate.Developing medium (such as cell culture medium 15) passes whole weighting material 3 with the form of plug flow.Once described vessel such as turn clockwise, developing medium (such as cell culture medium) just forces gas or air 16 displaced counter-clockwise at leading edge 20 place being in this plug flow.At trailing edge 21 place of medium 15, create optional limited low pressure, thus cause gas or air 16 to be aspirated by towards this trailing edge.Like this, medium 15 and gas or air 16 are through whole weighting material 3.

As a kind of replacement scheme shown in Fig. 2, the interior elongation wall 5 of alternative vessel 200 is provided by cylindrical tubular members 250.Identical Reference numeral instruction and similar characteristics disclosed in the vessel 1 about Fig. 1.

Outer tubular wall 4 is provided by outer tubular member 240.Interior elongation wall 5 is provided by elongated cylindrical tube element 250.Outer tubular member 240 and elongated cylindrical tube element 250 are fixed to two removable cappings 261 and 262.First capping 261 is provided with connection element 253, so that these vessel are attached to drive member, rotates to make the axis of these vessel along the longitudinal direction on 44.First capping 261 also comprises junctor 263, for internal space 2 is connected to conduit, and such as metal hose.

As an example, this outer tubular member can be Glass tubing, and it has the length L of such as 110mm and the internal diameter Do of such as 135mm.This interior elongate member can be poly(vinylidene fluoride) (PVDF) pipe, and it has the outer diameter D i of such as 88.9mm.Align with capping 261 and 262 in the outer end (therefore, the outer end of interior elongation wall) of this interior elongate member.These two cappings can be stainless steel or PVDF annular disc, and they can utilize silicone resin to be attached to described interior element and external component.First capping 261 can be provided with junctor 263, and the first capping 261 has connection element 253, and this connection element 253 has the outer diameter D ce of such as about 35mm.

Internal space 2 is filled with weighting material 3 at least partly.As an example, this weighting material can be identical or similar with the weighting material in the embodiment of Fig. 1.

Fig. 3 shows another alternative vessel 300.Identical Reference numeral instruction and similar characteristics disclosed in the vessel 1 about Fig. 1.

These vessel also comprise 2 fluid penetrable partition members 71 and 72, and described internal space is divided into 2 compartments 81 and 82 by this fluid penetrable partition member 71 and 72.Fluid penetrable partition member 71 and 72 extends to outer tubular wall 4 from interior elongation wall 5, and extends to the second capping 362 from the first capping 361 on the longitudinal direction 44 parallel with the direction of tubulose axis 45.A compartment 81 is provided with weighting material 3.Another compartment 82 is not provided with described weighting material.This fluid penetrable partition member can be provided with multiple hole body, such as, use porous material to provide porous partition member, or be provided with multiple hole, thus under any circumstance all allow liquid (i.e. developing medium) and gas to pass.But partition member 71 and 72 is provided with enough little hole body or hole, arrive opposite side to prevent this weighting material from the side of described partition member.

Outer tubular wall 4 is provided by outer tubular member 240, and outer tubular member 240 has the radial section along the plane vertical with longitudinal direction, and this cross section has round shape.

Interior elongation wall 5 is provided by interior elongate member 350, and it has along the radial section with the plane of vertical longitudinal direction, and this cross section has the shape of cutting circle or circular segment.In one embodiment, this circular segment has arc section 351 chord 352.Such as, the height of this circular segment has the size of 200mm (Dcc), 400mm (Do), 240mm (Di) and 125mm (L).In this embodiment, the string 352 of partition member 71 and 72 and this circular segment is coplanar.

The second capping 362 in two cappings 361 and 362 is provided with two junctors 364 and 365.First junctor 364 is arranged near outer tubular wall 4.Second junctor 364 is arranged near interior elongation wall 5.As shown in Figure 3, when these vessel have only been partially filled developing medium 1000, this medium has had liquid level 1001, and these vessel can rotate to as upper/lower positions: namely, and liquid does not contact interior elongation wall 5, but keeps given distance with interior elongation wall 5.When these vessel are in this position, the first junctor can be used for removing from compartment 82 or providing medium to compartment 82, and this compartment 82 is not filled with weighting material 3.Remove gas by junctor 364 from medium ullage or gas is provided to medium ullage.By making these vessel clockwise or be rotated counterclockwise, such as rotate maximum 360 ° or larger angle, medium through one of described two partition members, more particularly, will pass the partition member that will be submerged gradually in media as well.Along with described weighting material will due to rotation gradually through medium, medium will slowly through weighting material.Due to the rotation of these vessel, medium will pass with the form of plug flow and flow through whole weighting material.By the uniform contact between the medium that occurs in ring-type volume and weighting material.Once a part for described weighting material is through medium, medium just will ooze out gradually from described weighting material, and therefore, the gas in these vessel can contact described weighting material again, thus allows cell homoepitaxial on whole weighting material.

With reference to figure 4a and 4b, show another alternative vessel 400.The ring-type volume of internal space 2 is made up of multiple circular part 21,22,23 and 24 four/part of four ring-types (in this particular case, be).Each part 21,22,23 and 24 provides a part for outer tubular wall 4 respectively by outer tubular wall portion 441,442,443,444.Each part provides a part for interior elongation wall 5 respectively by interior elongation wall portion 451,452,453,454.

Each part has two radial part walls extended.As an example, part 21 has two radial part walls 4411 and 4412 extended.These part walls can transflective liquid and gas.Each part wall is provided with installation component 4413 and 4414, and this installation component 4413 and 4414 allows adjacent part to be coupled to each other.

First part's capping 461 and second section capping 462 limit in the first outer end of outer tubular wall and the second volume of outer end place to circular part and close respectively.The capping of these parts envelope forms the first capping and second capping of the ring-type volume 2 of these vessel together respectively.

As intelligible with further reference to Fig. 4 a-4b, each described part also can be provided with two fluid penetrable partition members, and the internal space of each circular part is divided into three compartments 481,482 and 483 by described two fluid penetrable partition members.Fluid penetrable partition member 471 and 472 (such as, porous partition member) extends to outer tubular wall 4 from interior elongation wall 5 and extends to the second capping 462 from the first capping 461 on the longitudinal direction 44 parallel with tubulose axis 405.A compartment 482 is provided with weighting material 3.Another two compartments 481 and 483 are not provided with described weighting material.These partition members can be similar or identical with the partition member of the vessel in Fig. 3.

Similar with shown in Fig. 3, is all arranged near outer tubular wall 4 for each circular part 21,22,23 and 24, first junctor 464.Second junctor 465 is arranged near interior elongation wall 5.Clearly visible from Fig. 4, when these vessel rotate around axis 405, each described part can use the separate vessel part of these vessel 400.Weighting material in each described part can be provided with medium, and this medium depends on the radial position of this part and is present in this part.

By installing multiple part (being four parts 21,22,23 and 24 in the present embodiment), obtain the vessel with outer tubular wall and interior elongation wall, the ring-type volume of these vessel is closed by two outer ends that two are sealed on outer tubular wall.Because the part walls extended by installing and connect two radial directions provide the connection between described multiple part, so collectively form of two sequential portion walls can the partition member of transflective liquid and gas, this partition member extends to outer tubular wall 4 from interior elongation wall 5 and extends to the second capping from the first capping on the longitudinal direction 44 parallel with the direction of tubulose axis 405.The present embodiment has the following advantages, that is: each part can be provided with the different media for different cell cultures.And, in the abnormal situation of reaction of the weighting material in one of described part, only can change a part.

As shown in this embodiment, the rotation of these vessel (or other vessel any disclosed herein) is provided by a turner.In one embodiment, this turner can comprise the outside surface contacting outer tubular wall with at least two supporting-point roller 490 with 491, and at least one in these at least two driving wheels is driven.

Fig. 4 c and 4d shows the alternate embodiments of vessel 401.In Fig. 4 c, the identical Reference numeral instruction part identical with Fig. 4 a and element.In the present embodiment, each part of vessel 401 has two radial part walls 4421 and 4422 extended.These part walls 4421 and 4422 can transflective liquid and gas.Each part wall 4421 comprises many holes 4423, and each in these holes exists corresponding hole in the second section wall 4422 of adjacent compartments.As shown in detail section B, the hole of first part's wall 4421 can be provided with outward extending edge 4425, and edge 4425 extends through the corresponding hole of second section wall 4422.Alternatively, the hole in adjacent portions bulkhead is provided with sealing member 4426, leaks between the wall contacted with each other to prevent medium.In alternative embodiments, between vessel, be provided with metal hose, instead of sealing member.

The embodiment of bio-reactor 500 has been shown in Fig. 5 a and Fig. 5 b.At least one vessel 501 similar with vessel 1 disclosed in Fig. 1 are rotatably arranged in container 502, container 502 can have any suitable radial section, the such as circle shown in Fig. 5 a, or Polygons, such as, rectangle (being chosen as square) shown in Fig. 5 b.This container has been partially filled developing medium 503, so that liquid level 504 is not increased to the axis 505 higher than this bio-reactor alternatively, but extends wall 509 at least contacting.Vessel 501 rotate around this axis 505, and this axis is equal to the longitudinal axis of vessel 501.The longitudinal axis of vessel 501 is axis parallel with the longitudinal direction of outer tubular wall 506.Alternatively, outer tubular wall 506 is provided with multiple hole, or is made up of porous material, such as liquids and gases permeable material.When the outer tubular wall 506 of this fluid penetrable rotate in the container 502 being partly filled with medium 503 make only outer tubular wall 506 a part of submergence in media as well time, can be weighting material 507 and the medium flowed into through outer tubular wall 506 in ring-type volume 508 be provided.When these vessel will rotate in described container, a part for medium can be pulled together with described weighting material.It will be appreciated by those skilled in the art that the further feature of setting forth for vessel 1,200,300 or 400 also can be added in vessel 501.

Another bio-reactor 600 has been shown in Fig. 6 and 6A.At least one vessel 601 (but in the figure, being two vessel 601 and 611 similar with vessel disclosed in Fig. 1) is rotatably arranged in container 602.This container has been partially filled developing medium 603, so that liquid level 604 is not increased to the axis 605 higher than this bio-reactor alternatively, but extends wall 609 at least contacting.Vessel 601 rotate around this axis 605, and axis 605 is equal to the longitudinal axis of vessel 601.The longitudinal axis of these vessel is axis parallel with the longitudinal direction of outer tubular wall.Alternatively, outer tubular wall 606 is provided with multiple hole, or is made up of porous material, such as liquids and gases permeable material.When this fluid penetrable outer tubular wall 606 rotate in the container 602 being partly filled with developing medium 603 part for only outer tubular wall 606 is immersed in developing medium 603 time, can be weighting material 607 and the developing medium 603 flowed into through outer tubular wall 606 in ring-type volume 608 be provided.When these vessel will rotate in described container, a part for developing medium 603 can be pulled together with described weighting material.

At least one (being vessel 601 in the present embodiment) in described vessel comprises magnetics 612.This bio-reactor also comprises magnetics 613, and magnetics 613 cooperates with the magnetics 612 of described vessel.Both magneticss 613 and 612 are positioned such that the magnetics 613 of bio-reactor 600 will cause the rotation of the magnetics 612 of vessel 601 around the rotation of rotation 605, and whole vessel 601 therefore will be made to rotate.Vessel adjacent one another are can be arranged on common axis 620, and they rotate around this common axis 620.Adjacent vessel 601 and 611 can be coupled to each other on fixed position, so that the rotation of vessel 601 also causes the rotation of vessel 611.

It will be understood by those skilled in the art that to there is the permeable outer tubular wall of liquids and gases but there is junctor and can install and rotate in the similar mode set forth about Fig. 5 and Fig. 6 with the vessel by optional metal hose these vessel being connected to gas or media reservoir.It will be understood by those skilled in the art that the further feature of setting forth for vessel 1,200,300 or 400 can be added in vessel 601.

As shown in Figure 7, vessel (vessel 300 of such as Fig. 3) can be rotated to provide bio-reactor by drive system.These vessel are arranged on rotatable spindle 701, and this rotatable spindle 701 can rotate around the rotation overlapped with the axis 45 of vessel 300.Axle 701 is set profile and is engaged on the unique position of rotation in the internal voids 702 of interior elongate member 350.Like this, by the position of rotation of Control Shaft 701, explicitly define the position of these vessel around axis 45.Described drive system also can comprise motor 703, such as linear motor, or other suitable device any, with the rotation of accurate Control Shaft 701.The trip bolt 704 preventing these vessel to be shifted along the longitudinal direction on axle 701 or any suitable device can fix these vessel 300 on axle 701 along the longitudinal direction 705 position.

Should be understood that alternatively, more than vessel 700 can be arranged on common axis 701.

Fig. 8 a-8c shows the cross-sectional view of the alternative example of the vessel be arranged on cooperation drive shaft 801.The shape of the shape of the internal voids 802 of interior elongate member 850 and the periphery of axle 801 is selected as this axle and vessel can be installed in the mode of uniqueness in a limited manner or even.

In the vessel 810 similar with the vessel of Fig. 1, in it, elongate member has the longitudinal fluting 811 in the inwall of this interior elongate member.Convex ridge 812 on axle 801 is engaged in this groove 811.These vessel coordinate on the shaft in a well defined manner.Convex ridge 812 can move in sliding manner in groove 811.

Vessel 820 are vessel similar with the vessel in Fig. 1.In it, elongate member has two longitudinal flutings 821 in the inwall of this interior elongate member.Two on axle 801 mutually perpendicular convex ridges 822 are engaged in these grooves 821.Vessel 820 are engaged on this axle in two ways, and first location rotates about the axis 180 ° relative to the second position.Convex ridge 822 can move in sliding manner in groove 821.

Vessel 830 are vessel similar with the vessel in Fig. 4 a and 4c.In it, elongate member has 4 longitudinal flutings 831, in each inwall of this interior elongate member provided by compartment, have one.Axle has the cross section of roughly cross-like, comprises 4 mutually perpendicular convex ridges 832 on axle 801.Each convex ridge is all engaged in the groove 831 of one of them compartment.Vessel 830 coordinate on the shaft in 4 kinds of modes.Convex ridge 832 can move in sliding manner in groove 831.

It will be understood by those skilled in the art that the further feature of the vessel described for Fig. 1 to 6 can be combined with drive system disclosed in Fig. 7 and 8.

With reference to figure 9, be shown as a part for bio-reactor 900 according to another alternative vessel 901 of the present disclosure.Identical Reference numeral indicates similar characteristics disclosed in the vessel 1 about Fig. 1.

The volume of internal space 2 is provided by multiple arcuate section 921,922,923 and 924 (in this particular case, being a 4 four/part).

Each described part provides a part for outer tubular wall 4 respectively by outer tubular wall portion 9441,9442,9443,9444.Each arch portion provides a part for interior elongation wall 5 respectively by interior elongation wall portion 9451,9452,9453,9454.

Each described arcuate section has two radial arcuate wall extended.As an example, arcuate section 921 has two radial arcuate wall 9411 and 9412 extended.These arcuate wall can transflective liquid and gas.

In the first outer end of outer tubular wall and the second outer end place, the volume 2 to arcuate section carries out gauge and closes respectively for first capping and the second capping, therefore closes the volume 2 of these vessel.

Each arcuate section also can be provided with two fluid penetrable partition members 9471 and 9472, and the internal space of each arc part is divided into 3 compartments 981,982 and 983 by these two fluid penetrable partition members 9471 and 9472.Fluid penetrable partition member 971 extends to outer tubular wall 4 and extends to the second capping from the first capping from interior elongation wall 5 with 972 on the longitudinal direction 944 parallel with tubulose axis 905 direction.One of them compartment 982 is provided with weighting material 3.Another two compartments 981 and 983 are not provided with described weighting material.These partition members can be similar or identical with the partition member of the vessel in Fig. 3.

Vessel 901 are rotatably arranged in container 900.This container has been partially filled developing medium 903, makes liquid level 904 not be increased to axis 905 higher than bio-reactor 900 alternatively.Vessel 901 rotate around this axis 905, and this axis 905 is equal to the longitudinal axis of vessel 901.The longitudinal axis of these vessel is axis parallel with the longitudinal direction of outer tubular wall.

Each compartment 981 and 983 is provided with multiple suitable hole, such as, respectively along longitudinal slit 991 of inwall 9451,9452,9453,9454.Each outer tubular wall portion 9441,9442,9443,9444 is respectively equipped with multiple hole or is made up of porous material, such as liquids and gases permeable material.When this fluid penetrable outer tubular wall rotate in the container 900 being partly filled with developing medium 903 part for only outer tubular wall is immersed in developing medium 903 time, the weighting material 3 that can be in described compartment provides the developing medium 903 flowed into through outer tubular wall in this volume.When these vessel will rotate in container, a part for developing medium 903 can be pulled together with described weighting material.Developing medium 903 also can fill the internal voids 9450 of interior elongation wall.Developing medium 903 can flow in part 981 and 982 or from part 981 and 982 through hole 991 and flow out, and alternatively, flows to adjacent part 982,981 respectively.

Figure 10 and 10a-10c shows the further example of vessel 1000, and these vessel 1000 comprise the main body 1002 that forms longitudinal wall and are the form of cap 1004,1006 in end.As shown in the figure, cap 1004,1006 can be removable, and in link position, form the liquid-tight seal being used for holding any fluid in main body 1002.Each cap 1004,1006 can comprise opening 1004a, 1006a, this 1004a, 1006a forms entrance or outlet (see Figure 11) for receiving developing medium, but also this entrance and exit all can be arranged in same cap, or be arranged in longitudinal wall of main body 1002.

Be provided with at least one fluid permeable structure (in shown situation, being a pair fluid permeable structure), such as perforated dividing plate 1008,1010, for the formation of the compartment 1012 (see Figure 12) holding any weighting material.Perforation in each dividing plate 1008 or 1010 can be set in shape and size, and to control the flowing of fluid in compartment 1012 and the residence time in compartment 1012, and these compartments can be same to each other or different to each other.

Described weighting material can comprise any one in said structure, but in one embodiment, can comprise fiber (such as woven or non-woven polyester fabric), especially nonwoven PET.Can this weighting material be set in the following manner, that is: makes it occupy the compartment of vessel 1000 completely, and therefore circumferentially contact internal surface and the described fluid permeable structure (such as dividing plate 1008,1010) of the wall of main body 1002.

Figure 11 shows vessel 1000 and can be associated with swivel arrangement 1100.Device 1100 can comprise pair of rolls 1102, for receiving, support vessel 1000 and making vessel 1000 rotate around the longitudinal axis of main body.In the case, vessel 1000 can be provided with substantial cylindrical main body 1002, this main body 1002 is suitable for engagement roller 1102 and is rotated by roller 1102, any fluid of any weighting material existed is distributed through in described compartment (such as with help, developing medium, or any clean-out system or restorative).

Tubular connector 1014,1016 also can be arranged to entrance 1004a and export 1006a and be associated, to deliver the fluid to described compartment.This can complete vessel 1000 are fixed while or while it rotates.In the case of the latter, junctor 1014,1016 can connect to allow the mode relatively rotated, such as, by using via the swivel joint of snap engagement generation or relatively rotating by using bearing.The sealing member of such as O shape ring can be used to help prevent any leakage and help to keep the sterile state desired by cell cultures.

An advantage of the vessel 1000 in Figure 10-12 is the simplicity of its structure.Such as, in an illustrated embodiment, vessel 1000 do not comprise sensor, probe, agitator etc.When wishing that it comprises these structures, it can comprise these structures, and as shown in figure 13, realizes by vessel 1000 and reservoir 1200 being connected with closed loop.Reservoir 1200 can comprise the sensor of any number, and to measure the one of the fluid be recycled or multiple characteristic, and it can comprise one-trip container (such as flexible pouch), to avoid necessity of cleaning and disinfection.Also pump can be provided with, such as peristaltic pump 1202, for making fluid via this circuit cycle.

The further aspect of the disclosure is described referring now to Figure 14-16.In this embodiment, can according to this vessel of any above-mentioned detail 1300 (thus forming a roller bottle), and these vessel 1300 comprise entrance 1302 and outlet 1304.Entrance 1302 and outlet 1304 all can be connected to conduit, at not excessively fixing (biding) or when reversing this conduit, this conduit allows vessel 1300 to rotate in the mode not disturbing fluid to transmit, and fluid thus can be allowed while these vessel rotate to flow into continuously in vessel 1300 and from vessel 1300 and flow out.Particularly, this conduit can comprise the flexible tubing 306 with open end, to be connected to entrance 1302 or outlet 1304 respectively, and can be associated with any fluid reservoirs at two end opposite places.Also a kind of suitable pumping equipment can be provided, for allowing fluid flow this conduit and vessel 1300.

Figure 15 and 16 shows the one possibility mode comprising the embodiment being applicable to the vessel allowing the conduit rotated used in Figure 14.Especially, as shown in figure 15, suitable turner (such as roller 1308) can be used to make vessel 1300 (such as clockwise) rotation one whole circle or more circle in a first direction.May but when not retraining this conduit, the described number of turns is variable, but it is expected to, 2-3 circle should be possible minimum value.Then, vessel 1300 can rotate whole circle or more circle in a second opposite direction.More particularly, this number of turns can be the number of turns in a first direction flexible tubing being returned to its original position, adds the corresponding number of turns in a second direction, as long as this flexible tubing does not retrain or otherwise disturb fluid to transmit.Above-mentioned rotation and reverse rotation can continuous the or occur discontinuously, and this is equally applicable to via the conveying of this conduit and reclaims fluid.

Later with reference to Figure 14, should also be understood that, when vessel 1300 comprise sensor 1310, this sensor 1310 also can be connected to any energy source for detecting.In this case, also can coil or be wound around any transmission line, such as cable 1312, to allow relative rotation when nothing is reversed or retrain in mode mentioned above.Again, it is desirable to any sensor to be arranged in any recirculation loop, because it reduce the cost of vessel 1300.

Other details of arrangement that Figure 16 and 17 shows the vessel 1400 of roller bottle form, that recommend.Such as, entrance 1402 and outlet 1404 can be arranged in common wall 1400a.Entrance 1402 also can be associated with the pipe of the fluid penetrable inner cylinder 1400b being positioned at fixed bed, and this contributes to guaranteeing that introduced fluid (gas, liquid or both) is not flow out immediately via entrance 1402 simply.

Figure 17 also shows by arranging injector 1420 (such as spinner-type flowmeter) in recirculation loop and is introduced in liquid by gas.This layout directly can be in the upstream of entrance 1402.As shown in figure 18, this gas incorporation way be associated with liquid-flow advantageously helps to reduce the appearance of foam and improves mass transfer velocity, because in gas is retained in by liquid separation air pocket (see Figure 19).Figure 18 also shows and exports 1404 transmission lines be associated 1422 and be back to reservoir 1424, ventilates by shown strainer 1426 pairs of reservoirs 1424, to avoid the ventilation hole for being directly connected with vessel 1400.

Should be understood that and can add other element to described vessel and bio-reactor.As an example, described interior elongation wall can be extend tubulose, is cylindrical wall alternatively.The internal voids of interior tubular wall can be used for holding one or more sensor, such as temperature sensor, position transducer are (such as, for limiting the direction of described vessel relative to rotation), optical pickocff (such as, for based on developing medium (such as cell culture medium) color produce data), pH sensor, oxygen sensor (such as dissolved oxygen (DO) sensor), CO ₂sensor, ammoniacal sensor or cell biological quantity sensor (such as, turbidity densometer).These sensors can additionally or alternatively be arranged in described capping or capping.

These vessel also can adapt to perfusion, continuously add fresh nutrient media and extract out and isopyknicly used medium, allow to realize as far as possible approx close to the cell culture condition of live body situation.Perfusion cell culture and the combination of such as enzymatic glucose biosensor allow the glucose consumption of monitoring continuously cell cultures.

Should also be understood that can be described outer wall alternatively for described inwall provides heating unit, such as heating cushion, for the temperature of the medium that heats or maintain in described vessel and weighting material.

Example

example 1: to the O_2 transport of weighting material material

With 66g carrier (packed bed of about 500ml) filled PVC cylindrical vessel (length 120mm, the internal diameter 75mm of nonwoven PET composition, external diameter 90mm), these vessel have outer wall, the inwall of perforation, two cappings, and one of them capping comprises cap.These vessel are filled with the water of 350ml.The described cap of these vessel is closed by the silicon cap (Siliconsen cultivates plug, flies the scientific and technological Bioblock of generation that) with sponge.Oxidative absorption dynamics data (completing measurement when probe is in liquid) is measured by the fluorescence DO probe be placed in these vessel (Presens company of Regensburg, Germany city).The cap of these vessel is unlocked, and nitrogen pours in vessel, until the complete desorb of oxygen (0%DO).Cultivate plug with Silicosen and close these vessel, and bottle is prevented on roller station.The sponge cultivating plug allows air to flow into.By measurement oxygen concentration along with the growth of time obtains oxygen absorption dynamics data.This experiment is performed with multiple different rotating speed.Under 2.60rpm, 5rpm and 6.65rpm, the DO in liquid rose to 75% (corresponding to 13h from 0% respectively in about 6 minutes, 5 minutes and 4 minutes ^-1, 16h ^-1, 38h ^-1k _i.a).

example 2: the small-scale Growth of Cells of the vessel be partially filled

Fill with the 1.33g carrier of nonwoven PET composition and there is 1350ml internal space volume and by cylindrical vessel (the long 120mm of 4 partition member gauges (limiting the angle of 90 degree), internal diameter 70mm, external diameter 130mm), comprise the packed bed part of 10ml, this cylindrical vessel has glass outer wall, the stainless steel inwall of perforation, a glass capping and a stainless steel capping containing cap.The cap of these vessel is cultivated plug by Siliconsen and is closed.These vessel through sterilizing and being filled with the developing medium (having the MEM of 1.8g/L glucose, the foetal calf serum of 5% and the non-essential amino acid of 1%) of 250ml, and can inoculate 12.2x10 ⁶mDBK cell (Ma Dashi bovine kidney cells) (i.e. 1.2x10 ⁶cell/cm ³packed bed).These vessel are placed on (6.5rpm) on roller, at 37 DEG C, are placed in culture dish.The gas supplied by described cap is enough to make the Growth of Cells duration of 7 days.In cultivation after 5 days, cell density reaches 18.2x10 ⁶cell/cm ³, and change medium.After 7 days, cell density reaches 46.0x10 ⁶cell/cm ³(that is, biomass increased 38.4 times in 7 days).

To be obvious for a person skilled in the art for realizing specifically implementing other arrangement of the target of vessel of the present invention.Should understand, although disclose preferred embodiment, ad hoc structure and structure and material for the various embodiments of device in this article, but not departing from the scope of the present invention be defined by the following claims, the various modifications and variations in form and details can be carried out.Such as, any formula given above all only represents available program.Function or therefrom delete function can be increased in block diagram, and, operation can be exchanged between functional blocks.Within the scope of the invention, can step be increased to described method or therefrom delete some steps.

Claims (96)

1. for vessel for cell cultures, described container has the inner compartment for Growth of Cells, and described vessel comprise:

-outer tubular wall, described outer tubular wall extends in a longitudinal direction, and described outer tubular wall delimit the outer edge of described inner compartment in radial directions;

-first end and the second end, described first end and the second end respectively at the first outer end of described outer tubular wall and the second outer end place to described inner compartment gauge; And

-weighting material, described weighting material to be in described inner compartment and to comprise fibre substrate.

2. vessel according to claim 1, further, wherein said first end and the second end comprise removable cap.

3. according to vessel according to claim 1 or claim 2, wherein, described first end comprises entrance, and described second end comprises outlet.

4. vessel according to claim 1, also comprise the one or more fluid penetrable dividing plates being positioned at described compartment, for contacting described fibre substrate.

5. a system, this system comprises the vessel according to any one in Claims 1-4 and the turner for rotating at least described weighting material.

6. system according to claim 5, wherein, described turner comprises the roller of the described outer tubular wall for contacting described vessel.

7. system according to claim 5, wherein, described turner comprises magnetic connecting parts.

8. the system according to any one in claim 5 to 8, also comprises closed loop, for delivering the fluid to described compartment and transferring out from described compartment.

9. system according to claim 8, also comprises the reservoir being connected to described closed loop.

10. system according to claim 9, also comprises at least one sensor be associated with described reservoir.

11. systems according to claim 10, wherein, described sensor is selected from the group be made up of following item: temperature sensor, position transducer, optical pickocff, pH sensor, oxygen sensor, CO ₂the combination of sensor, ammoniacal sensor, cell density sensor or any the sensor.

12. vessel according to any one in claim 1 to 11 or system, wherein, the fiber of described fibre substrate comprises nonwoven PET.

13. 1 kinds, for making the equipment of Growth of Cells, comprising:

-roller bottle, described roller bottle comprises: outer tubular wall, and described outer tubular wall extends in a longitudinal direction; First end and the second end, described first end and the second end carry out gauge in the first outer end of described outer tubular wall and the second outer end place to compartment respectively; And weighting material, described weighting material to be in described compartment and to comprise fibre substrate; With

-turner, described turner is for rotating described weighting material.

14. equipment according to claim 13, also comprise the first and second isolated first fluid permeable separator being positioned at described compartment.

15., according to claim 13 or equipment according to claim 14, also comprise closed loop, for delivering the fluid to described compartment and transferring out from described compartment.

16. equipment according to claim 15, also comprise the reservoir being connected to described closed loop.

17. equipment according to claim 16, also comprise at least one sensor be associated with described reservoir.

18. 1 kinds of vessel for cell cultures, described container has the inner compartment for Growth of Cells, and described vessel comprise:

-outer tubular wall, described outer tubular wall extends in a longitudinal direction, and described outer tubular wall delimit the outer edge of described inner compartment in the longitudinal direction;

-first end and the second end, described first end and the second end carry out gauge in the first outer end of described outer tubular wall and the second outer end place to described compartment respectively;

-weighting material, described weighting material is in described compartment; And

-at least one fluid penetrable dividing plate, at least one fluid penetrable dividing plate described is positioned at described compartment.

19. vessel according to claim 18, wherein, described dividing plate comprises perforation plate.

20. 1 kinds, for the method for culturing cell, comprising:

The weighting material comprising fibre substrate is provided to roller bottle;

Deliver the fluid in described roller bottle; With

Rotate described weighting material.

21. methods according to claim 20, wherein, the step rotating described weighting material rotates described roller bottle after being included in described supplying step.

22. methods according to claim 20, wherein, the step rotating described weighting material is included in described roller bottle and keeps fixed and rotate described weighting material simultaneously.

23. methods according to claim 20, extract the step of fluid out from described roller bottle during being also included in described spin step.

24. methods according to claim 20, detect the step of the characteristic of described fluid after being also included in described extraction step, and make extracted out fluid be back to step in described roller bottle during being included in described supplying step further.

25. 1 kinds of vessel for cell cultures, described container has the inner compartment for Growth of Cells, and described vessel comprise:

-outer tubular wall, described outer tubular wall extends in a longitudinal direction, and described outer tubular wall delimit the outer edge of described inner compartment in the longitudinal direction;

-first end and the second end, described first end and the second end carry out gauge in the first outer end of described outer tubular wall and the second outer end place to described compartment respectively;

-weighting material, described weighting material is in described inner compartment; And

-conduit, the fluid being used for culturing cell is transported in described inner compartment by described conduit.

26. vessel according to claim 25, wherein, described conduit comprises flexible tubing.

27. vessel according to claim 26, also comprise the entrance for being connected to described flexible tubing.

28. according to claim 26 or vessel according to claim 27, also comprise the outlet for being connected to described flexible tubing.

29. vessel according to claim 25, also include an inlet and an outlet.

30. vessel according to claim 29, wherein, described conduit comprises the first flexible tubing being connected to described entrance and the second flexible tubing being connected to described outlet.

31. vessel according to claim 29, wherein, described entrance and described outlet are arranged on the common wall of described vessel.

32. vessel according to claim 29, wherein, described entrance is connected to the pipe extended in described inner compartment, with at the position conveyance fluid away from described outlet.

33. vessel according to any one in claim 25 to 32, also comprise the sensor being connected to described vessel, described sensor is associated with flexible hose line.

34. vessel according to any one in claim 25 to 33, also comprise the insufflator being connected to described conduit.

35. vessel according to claim 34, wherein, described injector comprises spinner-type flowmeter.

36. vessel according to any one in claim 25 to 35, also comprise for the swivel joint with described tubes connection.

37. 1 kinds of systems for culturing cell, described system comprises the vessel according to any one in claim 25 to 36, these vessel be positioned at comprise described conduit recirculation loop on.

38. according to system according to claim 37, and wherein, described conduit is connected to described vessel at one end and is connected to reservoir at the other end place.

39. systems according to claim 35, wherein, described reservoir comprises strainer and/or sensor.

40. 1 kinds, for the method for culturing cell, comprising:

A () rotates the container with fixing weighting material along first direction; With

B (), after rotating along described first direction, rotates described container along second direction.

41. methods according to claim 40, comprising: during the step rotated along described first direction, deliver the fluid to described container.

42. methods according to claim 40, comprising: during the step rotated along described first direction, deliver the fluid to described container.

43. according to claim 41 or method according to claim 42, further comprising the steps of: before described supplying step, described container is connected to the conduit comprising flexible tubing.

44. methods according to any one in claim 41 to 43, wherein, described fluid comprises liquid, and described method is further comprising the steps of: before described supplying step, by gas injection in described liquid.

45. methods according to any one in claim 41 to 44, wherein, complete described rotation to the complete at least one times rotation on each direction of described container in described first direction and second direction.

46. methods according to any one in claim 41 to 44, wherein, complete described rotation at least twice complete rotation on each direction of described container in described first direction and second direction.

47. methods according to any one in claim 41 to 44, wherein, complete described rotation at least three complete rotations on each direction of described container in described first direction and second direction.

48. methods according to any one in claim 41 to 47, further comprising the steps of: repeating said steps (a) and (b).

49. 1 kinds of vessel for cell cultures, described vessel have annular internal space, and described annular internal space comprises the weighting material of applicable Growth of Cells, and described annular internal space has by with the ring-type volume of lower component gauge:

-outer tubular wall, longitudinal wall that described outer tubular wall has the first outer end and the second outer end and extends in a longitudinal direction, described outer tubular wall delimit the outer edge of described ring-type volume in the longitudinal direction;

-the first capping and the second capping, described first capping and the second capping also close described ring-type volume in the first outer end of described outer tubular wall and the second outer end place to described ring-type volume gauge respectively;

-Nei extends wall, first outer end of described interior elongation wall is directed by the first outer end towards described outer tubular wall, second outer end of described interior elongation wall is directed by the second outer end towards described outer tubular wall, described interior elongation wall is positioned at described outer tubular wall, described interior elongation wall extends in the longitudinal direction and delimit the inner boundary of described ring-type volume, and described inner boundary is surrounded by described outer edge.

50. vessel according to claim 49, wherein, tangent and the smallest circle cylinder with axis in a longitudinal direction has diameter Dmax, in the scope of the ultimate range radially between described inner boundary and outer edge between 5% to 45% of Dmax with described outer edge.

51. vessel according to any one in claim 49 or 50, wherein, tangent and the smallest circle cylinder with axis in a longitudinal direction has diameter Dmax, in the scope of the distance radially between described inner boundary and outer edge between 5% to 45% of Dmax with described outer edge.

52. vessel according to any one in claim 49 to 51, wherein, described weighting material is the weighting material of applicable adherent cell growth.

53. vessel according to any one in claim 49 to 51, wherein, described weighting material is the weighting material of applicable suspension culture.

54. vessel according to any one in claim 49 to 53, wherein, described weighting material is selected from the group be made up of following item: woven or nonwoven primitive fiber, porous carbon and glycan substrate.

55. vessel according to any one in claim 49 to 54, wherein, described vessel also comprise at least permeable partition member of first fluid and the permeable partition member of second fluid, described internal space is divided at least two compartments by the permeable partition member of described first fluid and the permeable partition member of second fluid, wherein:

-described fluid penetrable partition member extends to described outer tubular wall from described interior elongation wall and extends to described second capping from described first capping on the longitudinal direction parallel with the direction of tubulose axis, and

One in-described first compartment and the second compartment does not comprise weighting material.

56. vessel according to claim 55, wherein, the volume of the described compartment of non-filler is between 25% to 90% of the described ring-type volume of described internal space.

57. vessel according to any one in claim 55 to 56, wherein, described fluid penetrable partition member can transflective liquid and gas, and be suitable for keeping described weighting material.

58. vessel according to any one in claim 49 to 57, wherein, at least one in described first capping and the second capping is provided with at least one junctor, for providing medium and/or gas to described internal space and/or extract medium and/or gas from described internal space.

59. vessel according to claim 58, wherein, at least one in described first capping and the second capping comprises a more than junctor, for providing medium and/or gas to described internal space and/or extract medium and/or gas from described internal space.

60. vessel according to any one in claim 58 to 59, wherein, at least one in described first capping and the second capping is provided with at least one junctor, for providing medium and/or gas to described internal space and/or extract medium and/or gas from described internal space, at least one capping described comprising at least one junctor described is positioned such that at least one junctor described is applicable to not existing wherein in the described compartment of weighting material in the described compartment providing medium and/or gas and/or never there is weighting material and extracts medium and/or gas.

61. vessel according to any one in claim 58 to 60, wherein, at least one junctor described comprises the joint that closely cooperates for connecting flexible conduit.

62. vessel according to any one in claim 58 to 61, wherein, at least one junctor described is arranged in described capping near described outer tubular wall.

63. vessel according to any one in claim 49 to 62, wherein, described interior elongation wall is positioned at the center of described outer tubular wall.

64. vessel according to any one in claim 49 to 63, wherein, described outer tubular wall is provided by cylindrical tubular members.

65. vessel according to claim 63, wherein, described interior elongation wall is provided by cylindrical elements.

66. vessel according to claim 65, wherein, described interior elongate member is disposed coaxially in described outer tubular wall.

67. vessel according to any one in claim 65 to 66, wherein, described interior elongation wall has the radial section based on the plane vertical with the longitudinal direction of described interior elongation wall, and the shape of described radial section is selected from following group: circle, oval, ellipse, square, rectangle and circle are arc.

68. vessel according to claim 67, wherein, described interior elongation wall has the radial section based on the plane vertical with the longitudinal direction of described interior elongation wall, and described radial section has circular segment shape, and described circular segment has the central angle being greater than 180 °.

69. vessel according to any one in claim 49 to 68, wherein, described interior elongation wall is tubular wall.

70. vessel according to any one in claim 49 to 69, wherein, described interior elongation wall is connected in the fixed position for described outer tubular wall.

71. vessel according to claim 70, wherein, described interior elongation wall is provided by interior elongate member, and described outer tubular member is provided by outer tubular member, and described interior elongate member and described outer tubular member are by shaping integratedly.

72. vessel according to any one in claim 49 to 71, wherein, described outer tubular wall has the radial section based on the plane vertical with the longitudinal direction of described outer tubular wall, and described radial section has the shape selected from following group: circle, oval, ellipse, square or rectangle.

73. vessel according to any one in claim 49 to 72, wherein, described outer tubular wall and described interior elongation wall are cylindrical tubular wall, and in the scope of the radial width of the described internal space wherein, between described outer tubular wall and described interior elongation wall between 5% to 35% of the diameter of described outer tubular member.

74. vessel according to any one in claim 49 to 73, wherein, described interior elongation wall is disposed coaxially in described outer tubular wall.

75. vessel according to any one in claim 49 to 74, wherein, described internal space has the volume of 100 milliliters to 25 liters.

76. vessel according to any one in claim 49 to 75, wherein, the diameter of described outer tubular wall and the length ratio of described vessel are between 10/25 to 100/25.

77. vessel according to any one in claim 49 to 76, wherein, at least one in described first capping and the second capping comprises connection element, for described outer tubular wall being attached to the driver element for rotating described outer tubular wall.

78. vessel according to any one in claim 49 to 77, wherein, described interior elongation wall energy is enough to be provided by interior hollow elongated element, the first outer end of elongate member in described in described first lidstock.

79. vessel according to any one in claim 49 to 78, wherein, described interior elongation wall is provided by interior hollow elongated element, the second outer end of elongate member in described in described second lidstock.

80. vessel according to any one in claim 49 to 79, wherein, described interior elongation wall is fluid penetrable wall, and/or described outer tubular wall is fluid penetrable wall.

81. vessel according to any one in the claims, also comprise the vibrator for vibrating described vessel.

82. 1 kinds of bio-reactors for cell cultures, described bio-reactor comprises the one or more vessel according to any one in claim 49 to 80, wherein, described one or more vessel are configured such that the tubulose axis basic horizontal of described one or more vessel and can longitudinally rotate by axis.

83. bio-reactors according to Claim 8 described in 2, wherein, described one or more vessel comprise flexible conduit, for the internal space of described vessel being attached to the holder of medium and/or gas.

84. bio-reactors described in any one according to Claim 8 in 2 to 83, described bio-reactor comprises drive member, and described drive member can alternately clockwise and be rotated counterclockwise described outer tubular wall.

85. bio-reactors according to Claim 8 described in 4, described bio-reactor also comprises driving roll, and described driving roll is used for rotating described outer tubular wall by contacting the outside surface of described outer tubular wall.

86. 1 kinds of methods for culturing cell, described method comprises:

A) following material is incorporated in the vessel according to any one in claim 49 to 81:

-cell, and

-developing medium;

B) described vessel are located so that the longitudinal direction basic horizontal of described outer tubular wall, described outer tubular wall can rotate along the axis parallel with the longitudinal direction of described outer tubular wall; And

C) the described outer tubular wall of described vessel is made to rotate along described axis.

87. methods according to Claim 8 described in 6, wherein, described cell is attached cell, and is grown the described cultivation obtaining described cell by adherent cell.

88. methods according to Claim 8 described in 6, wherein, described cell is non-adherent cell, and obtains the cultivation of described cell by suspension culture.

89. methods described in any one according to Claim 8 in 6 to 88, wherein, when making described vessel be at least one radial position, the upper flow liquid level of described developing medium at least reaches described interior elongation wall.

90. methods described in any one according to Claim 8 in 6 to 89, wherein, when making described vessel be at least one radial position, the upper flow liquid level of described developing medium is lower than described interior elongation wall.

91. methods described in any one according to Claim 8 in 6 to 90, wherein, the volume of the medium arranged in open spaces is between 25% to 100% of the described ring-type volume of the internal space of described vessel.

92. methods described in any one according to Claim 8 in 6 to 91, wherein, make described outer tubular wall alternately clockwise and be rotated counterclockwise.

93. methods described in any one according to Claim 8 in 6 to 92, wherein, perform the rotation around described axis with the maximum angular rate between per minute 0.1 to 25 turn.

94. methods described in any one according to Claim 8 in 6 to 93, wherein, tubular wall has the outside surface with described internal space opposite orientation, and described outside surface has the maximum line velocity being up to 10cm/ second.

95. methods described in any one according to Claim 8 in 6 to 94, further comprising the steps of: to chemically examine and/or the medium changed in the described internal space of described vessel and/or gas.

96. methods described in any one according to Claim 8 in 6 to 95, go back the step of vessel described in involving vibrations.